利用Hoechst 33258荧光染色法检测紫外线B(UVB)辐射诱导HaCaT细胞凋亡率。结果表明,扇贝多肽(Polypeptide from Chlamys farreri,PCF)可以剂量依赖性抑制UVB诱导的HaCaT细胞凋亡;表皮生长因子受体(EGFR)抑制剂AG1478能明显抑制UVB诱导的...利用Hoechst 33258荧光染色法检测紫外线B(UVB)辐射诱导HaCaT细胞凋亡率。结果表明,扇贝多肽(Polypeptide from Chlamys farreri,PCF)可以剂量依赖性抑制UVB诱导的HaCaT细胞凋亡;表皮生长因子受体(EGFR)抑制剂AG1478能明显抑制UVB诱导的HaCaT细胞凋亡。采用3’-RACE法构建EGFR的cDNA片段,克隆测序检测突变位点。结果表明,UVB照射后EGFR发生碱基突变A→G,A→G,T→C,G→A,G→A;预加入5.69mmol/L的PCF,产生部分抗突变作用,1,2,3突变位点处未发生碱基的突变。展开更多
Biofouling is an important factor that affects bivalve farming industry. Fouling organisms may reduce growth and survival rate of the cultured species. Fouler are often filter feeders, so they are potential competitor...Biofouling is an important factor that affects bivalve farming industry. Fouling organisms may reduce growth and survival rate of the cultured species. Fouler are often filter feeders, so they are potential competitors for food resource with the cultured species. The present study was conducted to measure the impact of fouling on food uptake and nutrient release in April, 2006 in Daya Bay near Guangzhou, China. Results showed that fouling organisms had significant effect on food uptake and nutrient release. The chlorophyll a uptake rate of fouled scallops was 7.53 / Lh ±1.416 / Lh in April, significantly higher than those of cleaned scallops, Le., 4.23 / Lh±2.744 / Lh. The consumption of total particulate matter by fouled scallops in April was 5.52 / Lh± 0.818 / Lh; the corresponding results for cleaned scallops are 2.49 / Lh ±0.614 / Lh Fouling increased ammonia release significantly. The ammonia release rate of fouled scallops was 33.81/Lh ±7.699 / Lh in April, while cleaned scallops released 2.46 / Lh ±0.511 / Lh ammonia. Phosphate uptake of fouled scallops was 2.01± 0.699 / Lh and cleaned scallops released phosphate 6.01 / Lh ＋ 0.876 / Lh in April. There was not significant difference in nitrate consumption between fouled and cleaned scallops. According to the phytoplankton classification of input and output water samples, fouled scallops filtered more phytoplankton species than cleaned scallops. Therefore, this study showed that fouling contributed much to phytoplankton depletion and concentration increase of ammonia in water.展开更多
In April of 2006, a base population of the noble scallop Chlamys nobilis was established by collecting parental breeders from the stocks in Wushi, Zhanjiang. In December of 2006, 200 individuals were randomly sampled ...In April of 2006, a base population of the noble scallop Chlamys nobilis was established by collecting parental breeders from the stocks in Wushi, Zhanjiang. In December of 2006, 200 individuals were randomly sampled from the base population and subjected for correlation and path coefficient analysis. It was found that there were statistically significant phenotypic correlations among the traits （P 〈 0.01）. Total weight was significantly and positively correlated with the shell length （r = 0.934 3）, shell height （r = 0.895 9）, shell width （r = 0.899 1 ）, muscle weight （r = 0.882 0） and shell weight （r = 0.937 9）, respectively. Shell length, shell width, muscle weight, shell height and shell weight had positive and direct effects on the total weight, with values of 0.397 1, 0.321 9, 0.172 1, 0.089 6 and 0.066 9, respectively. Shell length, shell width and muscle weight had higher direct effects on the total weight than shell height and shell weight. A combined evaluation of correlation, direct effects and indirect effects showed that direct selection for shell length, shell width, muscle weight, shell height and shell weight would be effective to improving the total weight. It was concluded that these traits could be regarded as the selection criteria in breeding programs of the species.展开更多
Based on the research of juvenile (2, 3, 4 months) growth and survival of three populations of two different geographic areas in Chlamys farreri from Russian and China and their F1 hybrids derived from Chinese cultura...Based on the research of juvenile (2, 3, 4 months) growth and survival of three populations of two different geographic areas in Chlamys farreri from Russian and China and their F1 hybrids derived from Chinese cultural population (CC) (?) × Russian population (RW) (?) , Chinese wild population (CW) (?) × Russian population (RW) (?), Russian population (RW) (?) × Chinese wild population (CW) (?) , the study of the medium-term (6, 8, 10, 12 months) growth and development of Chlamys farreri was carried out. The four determined results indicated that there existed different extent heterosis (3% -52%) for the growth in three types of F1 hybrids, and the offspring derived from CC(?) ×R(?) had a stronger heterosis among the crosses at the medium-term; the uptrend among traits are wet weight >shell width>shell length> shell height, Chinese cultural population could be recognized as excellent parent, and seasonal variations influence very much on the daily increment and growth rate of each trait of Chlamys farreri and it is only able to survive and could barely grow in winter (6-8 months), but grows fast in temperate season (10-12 months).展开更多
The effects of 60 day starvation on survival rate, condition index (CI), changes of nutrient composition of different tissues, respiration and excretion of scallop Chlamys farreri were studied in laboratory from Oct. ...The effects of 60 day starvation on survival rate, condition index (CI), changes of nutrient composition of different tissues, respiration and excretion of scallop Chlamys farreri were studied in laboratory from Oct. 17 to Dec. 15,1997. Two groups (control and starvation with 200 individuals each) were cultured in two 2 m 3 tanks, with 31 to 32 salinity water at 17℃. Starvation effects were measured after 10, 20, 40 and 60 days. There was no mass mortality of scallops of the two tanks and survival rates of the control and starvation groups were 93.5% and 92.0%, respectively. Starvation had strong effect on the meat condition of the scallops, especially after 10 days; when relative lipid percentage dropped sharply while relative protein percentage increased. The impact of starvation on the oxygen consumption rate (OCR) and the ammonia N excretion rate (AER) was obvious. The OCR increased rapidly after 10 days but decreased after 20 days. The AER increased after 10 days and 20 days, but decreased obviously from 20 to 40 days. The O:N ratios varied to different degrees, and minimized after 20 days. The low O:N ratios implied that the protein was the main material for the metabolism of C. farreri.展开更多
Toll-like receptor(TLR) signaling pathway plays a pivotal role in the innate immune system. Studies on TLR signaling pathway genes in Zhikong scallop(Chlamys farreri) have mainly focused on sequence analysis and expre...Toll-like receptor(TLR) signaling pathway plays a pivotal role in the innate immune system. Studies on TLR signaling pathway genes in Zhikong scallop(Chlamys farreri) have mainly focused on sequence analysis and expression profiling, no research has been carried out on their localization. The chromosomal position of TLR signaling pathway genes can be valuable for assemblying scallop genome and analysizing gene regulatory networks. In the present study, five key TLR signaling pathway genes(Cf TLR, Cf Myd88, Cf TRAF6, Cf NFκB, and Cf IκB) containing bacterial artificial chromosomes(BACs) were isolated and physically mapped through fluorescence in situ hybridization on five non-homologous chromosome pairs, showing a similar distribution to another five model species. The isolation and mapping of these key immune genes of C. farreri will aid to the research on innate immunity, assignment of interested genes to chromosomes, and integration of physical, linkage and cytogenetic maps of this species.展开更多
Scallop is a popular sea food and an important aquaculture shellfish.Identification of genes and genetic variants relating to scallop growth could benefit high-yielding scallop breeding.Myostatin(MSTN) is a conservati...Scallop is a popular sea food and an important aquaculture shellfish.Identification of genes and genetic variants relating to scallop growth could benefit high-yielding scallop breeding.Myostatin(MSTN) is a conservative regulator of muscle growth,and has become one of the most important target genes for genetic improvement of the production of farmed animals.In this study,four single nucleotide polymorphisms(SNPs) were identified in the 5' flanking region of MSTN gene(Cf MSTN) in Zhikong scallop(Chlamys farreri).The association of these SNPs with scallop growth traits,including shell length,shell height,body weight and striated muscle weight was analyzed.The SNP g-1162G>T was found to associate with shell length,shell height,and striated muscle weight.The TT type scallops showed significantly higher trait values than those of GT type,and the GG type individuals exhibited median values.On the contrary,significantly more Cf MSTN transcripts were detected in the striated muscle of GT type scallops than in those of TT and GG type ones.Our results suggested that Cf MSTN might regulate the scallop muscle growth negatively,and SNP g-1162G>T can be used as a candidate marker for the selective breeding of high-yielding scallop.展开更多
DAX1, a member of nuclear receptor superfamily, has a function in the sex determination and gonadal differentiation of several vertebrate species. However, little information about DAX1 of invertebrates is available. ...DAX1, a member of nuclear receptor superfamily, has a function in the sex determination and gonadal differentiation of several vertebrate species. However, little information about DAX1 of invertebrates is available. Here we cloned a homolog of scallop(Chlamys farreri Jones and Preston 1904) dax1, Cf-dax1, and determined its expression characteristics at m RNA and protein levels. The c DNA sequence of Cf-dax1 was 2093 bp in length, including 1404 bp open reading frame(ORF) encoding 467 amino acids. Unlike those of vertebrates, no conserved LXXLL-related motif was found in the putative DNA binding region of Cf-DAX1. Fluorescence in situ hybridization showed that Cf-dax1 located on the short arm of a pair of subtelocentric chromosomes. Tissue distribution analysis using semi-quantitative RT-PCR revealed that Cf-dax1 expressed widely in adult scallop tissues, with the highest expression level found in adductor muscle, moderate level in mantle, gill and testis, and low level in kidney, ovary and hepatopancreas. The result of quantitative real-time PCR indicated that the expression of Cf-dax1 was significantly higher(P<0.05) in testis than in ovary at the same stage, showing a sex-dimorphic expression pattern. Furthermore, immunohistochemical detection found that Cf-DAX1 mainly located in spermatogonia and spermatocytes of testis and in oogonia and oocytes of ovary, implying that DAX1 may involve in gametogenesis of bivalves.展开更多
Biofouling is an important factor that affects the bivalve farming industry. Fouling organisms may reduce growth and survival rate of the cultured species. Fouler are often filter feeders, so they are potential compet...Biofouling is an important factor that affects the bivalve farming industry. Fouling organisms may reduce growth and survival rate of the cultured species. Fouler are often filter feeders, so they are potential competitors for food resource with the cul- tured species. The present study was conducted to measure the impact of fouling on food uptake and nutrient release in April and June, 2006 in Daya Bay near Guangzhou, China. Results showed that fouling organisms had significant effect on food uptake and nutrient release. The chlorophyll a uptake rate of fouled scallops was 7.53 Lh-1 ± 1.416 Lh-1 and 11.94 Lh-1± 2.497 Lh-1 in April and June, respectively, significantly higher than those of cleaned scallops, i.e., 4.23 Lh-1 ± 2.744 Lh-1 and 2.57 Lh-1 ± 1.832 Lh-1 respectively. The consumption of total particulate matter by fouled scallops in April and June was 5.52 Lh-1 ± 0.818 Lh-1 and 3.07 Lh-1 ± 0.971 Lh-1, respectively; the corresponding results for cleaned scallops are 2.49 Lh-1 ± 0.614 Lh-1 and 2.37 ± 1.214 Lh-1, respectively. Fouling in- creased ammonia release significantly. The ammonia release rate of fouled scallops was 33.81 Lh-1±7.699 Lh-1 and 76.39 Lh-1 ± 9.251 Lh-1 in April and June, while cleaned scallops released 2.46 Lh-1 ± 0.511 Lh-1 and 7.23 Lh-1 ± 1.026 Lh-1 ammonia, respectively. Phos- phate release of fouled scallops was 22.72 Lh-1 ± 9.978 Lh-1 in June and cleaned scallops released phosphate 6.01 Lh-1 ± 0.876 Lh-1 in April. Therefore, fouling contributed much to food reduction and concentration increase of ammonia and phosphate in water.展开更多
Polypeptide from Chlamys farreri (PCF) , a topical polypeptide isolated from Chlamys farreri , was used in this experiment aimed to investigate the photoprotective effect of PCF against chronic skin damage induced by ...Polypeptide from Chlamys farreri (PCF) , a topical polypeptide isolated from Chlamys farreri , was used in this experiment aimed to investigate the photoprotective effect of PCF against chronic skin damage induced by ultraviolet A (UVA) and ultraviolet B (UVB) radiation. The chronic ultraviolet irradiated guinea pig model was established, and visible changes in the skin including wrinkling, sagging and erythema were observed. Malondialdehyde (MDA) and antioxidant enzymes including superoxide dismutase (SOD) and glutathione peroxidase (GSH px) in the dorsal skin were determined using biochemical methods. The results showed:(1)PCF (5 % and 20%) could greatly protect the dorsal skin of guinea pig against wrinkling, sagging and erythema induced by UV radiation in a concentration dependent manner.(2)PCF could reduce MDA formation in the dorsal skin caused by UV irradiation, while increasing the activities of SOD and GSH px.(3)The differences among the PCF groups and UV model group were significant ( P <0.05, P <0.01). These results indicated that topical application of PCF provided broad solar UV spectrum photoprotection; and that the antioxidant property of PCF might play a role in photoprotection.展开更多
This study investigated the effect of a chitosan oligosaccharide-Ca complex (COS-Ca) on the depuration of cadmium (Cd) from Chlamys ferrari. After exposure to 0.5 mg L-1 CdCl2 for 3 or 7 d, the scallops were treated b...This study investigated the effect of a chitosan oligosaccharide-Ca complex (COS-Ca) on the depuration of cadmium (Cd) from Chlamys ferrari. After exposure to 0.5 mg L-1 CdCl2 for 3 or 7 d, the scallops were treated by COS-Ca prior to determina-tion of Cd, calcium (Ca) and zinc (Zn) contents, Cd distribution in organs, malondialdehyde (MDA) content and antioxidant variables. Results showed that COS-Ca reduced Cd content in the viscera of the scallops, with highest Cd depuration rate (47%) observed on day 3. The COS-Ca concentration substantially affected Cd depuration, and the exposure to 8.75 mg L-1 COS-Ca led to significantly higher Cd depuration rate compared with those of lower COS-Ca concentrations (1.75, 3.5, 5.25, and 7.00 mg L-1). Distribution analysis of Cd in scallop organs indicated that COS-Ca significantly reduced Cd content in the kidney throughout the 5-d experiment, as well as in the gill during the early stage of Cd depuration. In addition, COS-Ca treatment decreased glutathione peroxidase (GSH-Px) activity and MDA content while increasing superoxide dismutase (SOD) and catalase (CAT) activities on different days. Our work suggested COS-Ca complex treatment as an effective method for acceleration of Cd depuration from Cd-contaminated bivalves.展开更多
We investigated the radical scavenging effect and antioxidation property of polypeptide ex- tracted from Chlamys farreri (PCF) in vitro using chemiluminescence and electron spin resonance (ESR) methods. We examined th...We investigated the radical scavenging effect and antioxidation property of polypeptide ex- tracted from Chlamys farreri (PCF) in vitro using chemiluminescence and electron spin resonance (ESR) methods. We examined the scavenging effects of PCF on superoxide anions ( O?2 ), hydroxyl radicals (OH·), peroxynitrite (ONOO-) and the inhibiting capacity of PCF on peroxidation of linoleic acid. Our experiment suggested that PCF could scavenge oxygen free radicals including superoxide anions ( O2? ) (IC50 =0.3 mg/ml), hydroxyl radicals (OH·) (IC50 = 0.2 μg/ml) generated from the reaction systems and effectively inhibit the oxidative activity of ONOO- (IC50 = 0.2 mg/ml). At 1.25 mg/ml of PCF, the inhibi- tion ratio on lipid peroxidation of linoleic acid was 43 %. The scavenging effect of PCF on O?2 , OH· and ONOO- free radicals were stronger than those of vitamin C but less on lipid peroxidation of linoleic acid. Thus PCF could scavenge free radicals and inhibit the peroxidation of linoleic acid in vitro. It is an anti- oxidant from marine products and potential for industrial production in future.展开更多
This study applied an optimized one-step 2b-RAD library construction strategy and performed simplified genome sequencing of 539 individuals from three continuously selected Chlamys farreri populations. SNP screening w...This study applied an optimized one-step 2b-RAD library construction strategy and performed simplified genome sequencing of 539 individuals from three continuously selected Chlamys farreri populations. SNP screening was performed using RAD typing software and population genetic parameters for the continuously selected populations from three generations(G1, G2, G3) were determined. The results showed that the optimized one-step 2b-RAD library construction strategy greatly simplified the experimental process, making it suitable for efficiently constructing a large number of libraries. A total of 18450 SNP markers were identified, which evenly distributed throughout the genome. Population genetic analysis of these three generations showed that the mean value of observed heterozygosity was 0.275 ± 0.177, 0.272 ± 0.181 and 0.275 ± 0.166, respectively. Meanwhile, the mean value of expected heterozygosity was 0.275 ± 0.141, 0.274 ± 0.145 and 0.280 ± 0.133, respectively. The Wright's fixation index(F) was 0.04291, 0.04976 and 0.06685, respectively. Markers deviated from Hardy–Weinberg equilibrium accounted for 10.34%, 12.64%, and 23.11%, and the Shannon diversity index was 0.0999 ± 0.0404, 0.0921 ± 0.0388 and 0.0733 ± 0.0308. FIS(also known as the inbreeding coefficient) of the three populations was 0.0256, 0.0323 and 0.0468, respectively. We suggested that the 2b-RAD method is well suited to population genetic studies of aquacultured organisms. Moreover, our results indicated that the continuous selection affected the population genetic structure of the cultured Penglai-Red scallop, but the change was not significant; therefore, population selection should continue.展开更多
The biomass and size fraction of phytoplankton in terms of chlorophyll a（Chl a） was measured during four cruises conducted in April, July, October 2013 and January 2014 in mariculture area, the Sanggou Bay, China.Re...The biomass and size fraction of phytoplankton in terms of chlorophyll a（Chl a） was measured during four cruises conducted in April, July, October 2013 and January 2014 in mariculture area, the Sanggou Bay, China.Results show that total Chl a levels in the surface seawater of the Sanggou Bay generally range from 0.10 to 20.46μg/L, with an average value of 2.13 μg/L. Nano-phytoplankton was the most important size-fraction and accounted for about 65.1% of total Chl a. In order to evaluate the importance of the ＂protozoan trophic link＂ for energy transfer from the microbial loop to filter-feeding feeders, Zhikong scallop Chlamys farreri was then offered a natural planktonic community as potential prey. Results show that scallops obtained carbon source from natural plankton with the rate of 11 033.05 μg/（g·d）. Protists（nanoflagellates and ciliates） were the dominant source of carbon retained by scallop（48.78%）. The microbial loop provided 58.45% of the carbon source for farmed scallops. These results indicate that the microbial loop represent a valuable trophic resource in mariculture system of the Sanggou Bay.展开更多
Bivalve live in aquatic environment and the water temperature can affect their immunity directly.In this research,the scallop Chlamys farreri was injected with 10^4 or 10^7 CFU mL^-1 Vibrio anguillarum and cultured at...Bivalve live in aquatic environment and the water temperature can affect their immunity directly.In this research,the scallop Chlamys farreri was injected with 10^4 or 10^7 CFU mL^-1 Vibrio anguillarum and cultured at 11℃,17℃,23℃,and 28℃,respectively.For the control scallop,only phosphate-buffered saline(PBS)was injected.Then total haemocytes and granulocytes were measured by ELISA using monoclonal antibodies.In the meantime,expressions of six immunity-related genes,including lipopolysaccharide andβ-1,3-glucan binding protein(CfLGBP),C-type lectin(CfLec-2),Toll-like receptor(Cf TLR),Lysozyme(CfLYZ),superoxide dismutase(SOD),and phenoloxidase(CfPO)in haemocytes were measured using quantitative real-time PCR.The results showed that total haemocytes counts in 10^4 CFU mL^-1 injection groups showed no differences compared to the control group at all temperatures.However,they varied significantly in 10^7 CFU mL^-1 injection groups at 3 h at 11℃,6–12 h at 17℃,3–48 h at 23℃,and 12–48 h at 28℃.Granulocytes counts in 10^4 CFU mL^-1 injection groups showed no variance compared to the control group at all temperatures,except for 12 h at 23℃,and 24–36 h at 28℃.They were significantly decreased in 10^7 CFU mL^-1 injection groups during 6–48 h at 11℃,12–48 h at 17℃,3–48 h at 23℃,and 3–72 h at 28℃.The expression levels of six immunity-related genes in haemocytes of 10^7 CFU mL^-1 injection groups were significantly higher than those of control group and 10^4 CFU mL^-1 injection groups at all temperatures.The results indicated that infected with high concentration of vibrios,haemocyte counts,granulocyte counts and the expressions of immunity-related genes in scallop C.farreri were significantly affected by environmental temperature.展开更多
The effects of membrane penetrating anti-freezing agents(MPAAs), DMSO(dimethyl sulfoxide),glycerol,EG(ethylene glycol) and methanol in combination with different cryoprotective additives suchas carbohydrates,macromole...The effects of membrane penetrating anti-freezing agents(MPAAs), DMSO(dimethyl sulfoxide),glycerol,EG(ethylene glycol) and methanol in combination with different cryoprotective additives suchas carbohydrates,macromolecules and inorganic compounds on the spermatozoon vitality of Chinesescallop,Chlamys farreri,during 1 h 0℃ equilibrium were investigated.When only MPAAs existed,the detrimental effects of different MPAAs ranked in the following order:DMSO【methanol【EG【glycerol.When carbohydrates were added into MPAAs solution,5% glucose caused larger decrease ofspermatozoon vitality than 2.4% lactose.5% glucose or 2.4% lactose in 7.5% glycerol caused com-plete damage.10% yolk was best in maintaining the spermatozoon vitality except when used incombination with 10% methanol.10% milk significantly decreased spermatozoon vitality in EG andmethanol and enhanced its vitality in glycerol,but did not significantly influence it in DMSO.Glycinewas apparently detrimental to spermatozoon vitality.The average展开更多
Chlamys farreri is an economically important mollusk that can accumulate excessive amounts of cadmium(Cd). Studying the molecular mechanism of Cd accumulation in bivalves is difficult because of the lack of genome bac...Chlamys farreri is an economically important mollusk that can accumulate excessive amounts of cadmium(Cd). Studying the molecular mechanism of Cd accumulation in bivalves is difficult because of the lack of genome background. Transcriptomic analysis based on high-throughput RNA sequencing has been shown to be an efficient and powerful method for the discovery of relevant genes in non-model and genome reference-free organisms. Here, we constructed two c DNA libraries(control and Cd exposure groups) from the digestive gland of C. farreri and compared the transcriptomic data between them. A total of 227 673 transcripts were assembled into 105 071 unigenes, most of which shared high similarity with sequences in the NCBI non-redundant protein database. For functional classification, 24 493 unigenes were assigned to Gene Ontology terms. Additionally, Eu Karyotic Ortholog Groups and Kyoto Encyclopedia of Genes and Genomes analyses assigned 12 028 unigenes to 26 categories and 7 849 unigenes to five pathways, respectively. Comparative transcriptomics analysis identified 3 800 unigenes that were differentially expressed in the Cd-treated group compared with the control group. Among them, genes associated with heavy metal accumulation were screened, including metallothionein, divalent metal transporter, and metal tolerance protein. The functional genes and predicted pathways identified in our study will contribute to a better understanding of the metabolic and immune system in the digestive gland of C. farreri. In addition, the transcriptomic data will provide a comprehensive resource that may contribute to the understanding of molecular mechanisms that respond to marine pollutants in bivalves.展开更多
To investigate the effect of polypeptide from Chlamys farreri (PCF) on NHDF in vitro, we modeled oxidative damage on normal human dermal fibroblasts (NHDF) exposed to ultraviolet B (UVB). In this study, 3-[4,5-Dimethy...To investigate the effect of polypeptide from Chlamys farreri (PCF) on NHDF in vitro, we modeled oxidative damage on normal human dermal fibroblasts (NHDF) exposed to ultraviolet B (UVB). In this study, 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydro-genase (LDH) were tested to measure cell viability. Enzymes including superoxide dismutase (SOD), glu-tathione peroxidase (GSH-PX), catalase (CAT) and xanthine oxidase (XOD) were determined biochemically. Total antioxidative capacity (T-AOC) and anti-superoxide anion capacity (A-SAC) were also determined. Ultrastructure of fibroblasts was observed under transmission electron microscope. The results showed that: UVB (1.176×10-4 J/cm2) suppressed the growth of fibroblasts and the introduction of PCF (0.25%-l%) before UVB reduced the suppression in a concentration-dependent manner. PCF could enhance the activities of SOD, GSH-PX and T-AOC as well as A-SAC. Also PCF could inhibit XOD activity, while it did not affect CAT activity. Ultrastructure of fibroblasts were damaged after UVB irradiation, concentration-dependent PCF reduced the destructive effect of UVB on cells. These results indicated that PCF can protect human dermal fibroblasts from being harmed by UVB irradiation via its antioxidant pro-erty.展开更多
文摘利用Hoechst 33258荧光染色法检测紫外线B(UVB)辐射诱导HaCaT细胞凋亡率。结果表明,扇贝多肽(Polypeptide from Chlamys farreri,PCF)可以剂量依赖性抑制UVB诱导的HaCaT细胞凋亡;表皮生长因子受体(EGFR)抑制剂AG1478能明显抑制UVB诱导的HaCaT细胞凋亡。采用3’-RACE法构建EGFR的cDNA片段,克隆测序检测突变位点。结果表明,UVB照射后EGFR发生碱基突变A→G,A→G,T→C,G→A,G→A;预加入5.69mmol/L的PCF,产生部分抗突变作用,1,2,3突变位点处未发生碱基的突变。
基金supported by National Science Foundation of Huaihai Institute of Technology (No. KQ07102)Open Project Program of the Key Laboratory of Marine Bio-resources Sustainable Utilization, SCSIO, CAS (No. KK09001)
文摘Biofouling is an important factor that affects bivalve farming industry. Fouling organisms may reduce growth and survival rate of the cultured species. Fouler are often filter feeders, so they are potential competitors for food resource with the cultured species. The present study was conducted to measure the impact of fouling on food uptake and nutrient release in April, 2006 in Daya Bay near Guangzhou, China. Results showed that fouling organisms had significant effect on food uptake and nutrient release. The chlorophyll a uptake rate of fouled scallops was 7.53 / Lh ±1.416 / Lh in April, significantly higher than those of cleaned scallops, Le., 4.23 / Lh±2.744 / Lh. The consumption of total particulate matter by fouled scallops in April was 5.52 / Lh± 0.818 / Lh; the corresponding results for cleaned scallops are 2.49 / Lh ±0.614 / Lh Fouling increased ammonia release significantly. The ammonia release rate of fouled scallops was 33.81/Lh ±7.699 / Lh in April, while cleaned scallops released 2.46 / Lh ±0.511 / Lh ammonia. Phosphate uptake of fouled scallops was 2.01± 0.699 / Lh and cleaned scallops released phosphate 6.01 / Lh ＋ 0.876 / Lh in April. There was not significant difference in nitrate consumption between fouled and cleaned scallops. According to the phytoplankton classification of input and output water samples, fouled scallops filtered more phytoplankton species than cleaned scallops. Therefore, this study showed that fouling contributed much to phytoplankton depletion and concentration increase of ammonia in water.
文摘In April of 2006, a base population of the noble scallop Chlamys nobilis was established by collecting parental breeders from the stocks in Wushi, Zhanjiang. In December of 2006, 200 individuals were randomly sampled from the base population and subjected for correlation and path coefficient analysis. It was found that there were statistically significant phenotypic correlations among the traits （P 〈 0.01）. Total weight was significantly and positively correlated with the shell length （r = 0.934 3）, shell height （r = 0.895 9）, shell width （r = 0.899 1 ）, muscle weight （r = 0.882 0） and shell weight （r = 0.937 9）, respectively. Shell length, shell width, muscle weight, shell height and shell weight had positive and direct effects on the total weight, with values of 0.397 1, 0.321 9, 0.172 1, 0.089 6 and 0.066 9, respectively. Shell length, shell width and muscle weight had higher direct effects on the total weight than shell height and shell weight. A combined evaluation of correlation, direct effects and indirect effects showed that direct selection for shell length, shell width, muscle weight, shell height and shell weight would be effective to improving the total weight. It was concluded that these traits could be regarded as the selection criteria in breeding programs of the species.
基金This study is contribution No. G199901209 of 973 from the Chinese Basic Research Project and Chinese Postdoctor Found
文摘Based on the research of juvenile (2, 3, 4 months) growth and survival of three populations of two different geographic areas in Chlamys farreri from Russian and China and their F1 hybrids derived from Chinese cultural population (CC) (?) × Russian population (RW) (?) , Chinese wild population (CW) (?) × Russian population (RW) (?), Russian population (RW) (?) × Chinese wild population (CW) (?) , the study of the medium-term (6, 8, 10, 12 months) growth and development of Chlamys farreri was carried out. The four determined results indicated that there existed different extent heterosis (3% -52%) for the growth in three types of F1 hybrids, and the offspring derived from CC(?) ×R(?) had a stronger heterosis among the crosses at the medium-term; the uptrend among traits are wet weight >shell width>shell length> shell height, Chinese cultural population could be recognized as excellent parent, and seasonal variations influence very much on the daily increment and growth rate of each trait of Chlamys farreri and it is only able to survive and could barely grow in winter (6-8 months), but grows fast in temperate season (10-12 months).
文摘The effects of 60 day starvation on survival rate, condition index (CI), changes of nutrient composition of different tissues, respiration and excretion of scallop Chlamys farreri were studied in laboratory from Oct. 17 to Dec. 15,1997. Two groups (control and starvation with 200 individuals each) were cultured in two 2 m 3 tanks, with 31 to 32 salinity water at 17℃. Starvation effects were measured after 10, 20, 40 and 60 days. There was no mass mortality of scallops of the two tanks and survival rates of the control and starvation groups were 93.5% and 92.0%, respectively. Starvation had strong effect on the meat condition of the scallops, especially after 10 days; when relative lipid percentage dropped sharply while relative protein percentage increased. The impact of starvation on the oxygen consumption rate (OCR) and the ammonia N excretion rate (AER) was obvious. The OCR increased rapidly after 10 days but decreased after 20 days. The AER increased after 10 days and 20 days, but decreased obviously from 20 to 40 days. The O:N ratios varied to different degrees, and minimized after 20 days. The low O:N ratios implied that the protein was the main material for the metabolism of C. farreri.
基金financially supported by the National Natural Science Foundation of China(31270047)the National High Tech R&D Program(2012AA10A410)+1 种基金the National Basic Research Program of China(2010CB126402)the National Key Technology R&D Program of China(2011BAD45B01 and 2011BAD13B05)
文摘Toll-like receptor(TLR) signaling pathway plays a pivotal role in the innate immune system. Studies on TLR signaling pathway genes in Zhikong scallop(Chlamys farreri) have mainly focused on sequence analysis and expression profiling, no research has been carried out on their localization. The chromosomal position of TLR signaling pathway genes can be valuable for assemblying scallop genome and analysizing gene regulatory networks. In the present study, five key TLR signaling pathway genes(Cf TLR, Cf Myd88, Cf TRAF6, Cf NFκB, and Cf IκB) containing bacterial artificial chromosomes(BACs) were isolated and physically mapped through fluorescence in situ hybridization on five non-homologous chromosome pairs, showing a similar distribution to another five model species. The isolation and mapping of these key immune genes of C. farreri will aid to the research on innate immunity, assignment of interested genes to chromosomes, and integration of physical, linkage and cytogenetic maps of this species.
基金supported by the National High-Tech R&D Program (863 Program,2012AA10A402)Doctoral Fund of Ministry of Education of China (20120132130 002)the National Key Technology R&D Program of China (2011BAD13B06)
文摘Scallop is a popular sea food and an important aquaculture shellfish.Identification of genes and genetic variants relating to scallop growth could benefit high-yielding scallop breeding.Myostatin(MSTN) is a conservative regulator of muscle growth,and has become one of the most important target genes for genetic improvement of the production of farmed animals.In this study,four single nucleotide polymorphisms(SNPs) were identified in the 5' flanking region of MSTN gene(Cf MSTN) in Zhikong scallop(Chlamys farreri).The association of these SNPs with scallop growth traits,including shell length,shell height,body weight and striated muscle weight was analyzed.The SNP g-1162G>T was found to associate with shell length,shell height,and striated muscle weight.The TT type scallops showed significantly higher trait values than those of GT type,and the GG type individuals exhibited median values.On the contrary,significantly more Cf MSTN transcripts were detected in the striated muscle of GT type scallops than in those of TT and GG type ones.Our results suggested that Cf MSTN might regulate the scallop muscle growth negatively,and SNP g-1162G>T can be used as a candidate marker for the selective breeding of high-yielding scallop.
基金supported by the National High Technology Research and Development Program of China (863 Program) (2012AA10A402)the Natural Science Foundation of Qingdao (11-2-4-1(10)-jch)the Key Natural Science Foundation of Shandong Province (Z2008D02)
文摘DAX1, a member of nuclear receptor superfamily, has a function in the sex determination and gonadal differentiation of several vertebrate species. However, little information about DAX1 of invertebrates is available. Here we cloned a homolog of scallop(Chlamys farreri Jones and Preston 1904) dax1, Cf-dax1, and determined its expression characteristics at m RNA and protein levels. The c DNA sequence of Cf-dax1 was 2093 bp in length, including 1404 bp open reading frame(ORF) encoding 467 amino acids. Unlike those of vertebrates, no conserved LXXLL-related motif was found in the putative DNA binding region of Cf-DAX1. Fluorescence in situ hybridization showed that Cf-dax1 located on the short arm of a pair of subtelocentric chromosomes. Tissue distribution analysis using semi-quantitative RT-PCR revealed that Cf-dax1 expressed widely in adult scallop tissues, with the highest expression level found in adductor muscle, moderate level in mantle, gill and testis, and low level in kidney, ovary and hepatopancreas. The result of quantitative real-time PCR indicated that the expression of Cf-dax1 was significantly higher(P<0.05) in testis than in ovary at the same stage, showing a sex-dimorphic expression pattern. Furthermore, immunohistochemical detection found that Cf-DAX1 mainly located in spermatogonia and spermatocytes of testis and in oogonia and oocytes of ovary, implying that DAX1 may involve in gametogenesis of bivalves.
基金supported by National Science Foundation of Huaihai Institute of Technology(No.KQ07102)National Science Foundation of China(Grant Nos.30670401 and 40576052).
文摘Biofouling is an important factor that affects the bivalve farming industry. Fouling organisms may reduce growth and survival rate of the cultured species. Fouler are often filter feeders, so they are potential competitors for food resource with the cul- tured species. The present study was conducted to measure the impact of fouling on food uptake and nutrient release in April and June, 2006 in Daya Bay near Guangzhou, China. Results showed that fouling organisms had significant effect on food uptake and nutrient release. The chlorophyll a uptake rate of fouled scallops was 7.53 Lh-1 ± 1.416 Lh-1 and 11.94 Lh-1± 2.497 Lh-1 in April and June, respectively, significantly higher than those of cleaned scallops, i.e., 4.23 Lh-1 ± 2.744 Lh-1 and 2.57 Lh-1 ± 1.832 Lh-1 respectively. The consumption of total particulate matter by fouled scallops in April and June was 5.52 Lh-1 ± 0.818 Lh-1 and 3.07 Lh-1 ± 0.971 Lh-1, respectively; the corresponding results for cleaned scallops are 2.49 Lh-1 ± 0.614 Lh-1 and 2.37 ± 1.214 Lh-1, respectively. Fouling in- creased ammonia release significantly. The ammonia release rate of fouled scallops was 33.81 Lh-1±7.699 Lh-1 and 76.39 Lh-1 ± 9.251 Lh-1 in April and June, while cleaned scallops released 2.46 Lh-1 ± 0.511 Lh-1 and 7.23 Lh-1 ± 1.026 Lh-1 ammonia, respectively. Phos- phate release of fouled scallops was 22.72 Lh-1 ± 9.978 Lh-1 in June and cleaned scallops released phosphate 6.01 Lh-1 ± 0.876 Lh-1 in April. Therefore, fouling contributed much to food reduction and concentration increase of ammonia and phosphate in water.
文摘Polypeptide from Chlamys farreri (PCF) , a topical polypeptide isolated from Chlamys farreri , was used in this experiment aimed to investigate the photoprotective effect of PCF against chronic skin damage induced by ultraviolet A (UVA) and ultraviolet B (UVB) radiation. The chronic ultraviolet irradiated guinea pig model was established, and visible changes in the skin including wrinkling, sagging and erythema were observed. Malondialdehyde (MDA) and antioxidant enzymes including superoxide dismutase (SOD) and glutathione peroxidase (GSH px) in the dorsal skin were determined using biochemical methods. The results showed:(1)PCF (5 % and 20%) could greatly protect the dorsal skin of guinea pig against wrinkling, sagging and erythema induced by UV radiation in a concentration dependent manner.(2)PCF could reduce MDA formation in the dorsal skin caused by UV irradiation, while increasing the activities of SOD and GSH px.(3)The differences among the PCF groups and UV model group were significant ( P <0.05, P <0.01). These results indicated that topical application of PCF provided broad solar UV spectrum photoprotection; and that the antioxidant property of PCF might play a role in photoprotection.
基金supported by grants of the National Key Technology Research and Development Program in the 11th Five-Year Plan of China (2008BAD94B\09)the National Natural Science Foundation of China (Grant No. 30972289)
文摘This study investigated the effect of a chitosan oligosaccharide-Ca complex (COS-Ca) on the depuration of cadmium (Cd) from Chlamys ferrari. After exposure to 0.5 mg L-1 CdCl2 for 3 or 7 d, the scallops were treated by COS-Ca prior to determina-tion of Cd, calcium (Ca) and zinc (Zn) contents, Cd distribution in organs, malondialdehyde (MDA) content and antioxidant variables. Results showed that COS-Ca reduced Cd content in the viscera of the scallops, with highest Cd depuration rate (47%) observed on day 3. The COS-Ca concentration substantially affected Cd depuration, and the exposure to 8.75 mg L-1 COS-Ca led to significantly higher Cd depuration rate compared with those of lower COS-Ca concentrations (1.75, 3.5, 5.25, and 7.00 mg L-1). Distribution analysis of Cd in scallop organs indicated that COS-Ca significantly reduced Cd content in the kidney throughout the 5-d experiment, as well as in the gill during the early stage of Cd depuration. In addition, COS-Ca treatment decreased glutathione peroxidase (GSH-Px) activity and MDA content while increasing superoxide dismutase (SOD) and catalase (CAT) activities on different days. Our work suggested COS-Ca complex treatment as an effective method for acceleration of Cd depuration from Cd-contaminated bivalves.
基金Supported by the National Natural Science Foundation of China (No. 39970638) and the Science and Technology Bureau of Qingdao (No. 2001-28-50).
文摘We investigated the radical scavenging effect and antioxidation property of polypeptide ex- tracted from Chlamys farreri (PCF) in vitro using chemiluminescence and electron spin resonance (ESR) methods. We examined the scavenging effects of PCF on superoxide anions ( O?2 ), hydroxyl radicals (OH·), peroxynitrite (ONOO-) and the inhibiting capacity of PCF on peroxidation of linoleic acid. Our experiment suggested that PCF could scavenge oxygen free radicals including superoxide anions ( O2? ) (IC50 =0.3 mg/ml), hydroxyl radicals (OH·) (IC50 = 0.2 μg/ml) generated from the reaction systems and effectively inhibit the oxidative activity of ONOO- (IC50 = 0.2 mg/ml). At 1.25 mg/ml of PCF, the inhibi- tion ratio on lipid peroxidation of linoleic acid was 43 %. The scavenging effect of PCF on O?2 , OH· and ONOO- free radicals were stronger than those of vitamin C but less on lipid peroxidation of linoleic acid. Thus PCF could scavenge free radicals and inhibit the peroxidation of linoleic acid in vitro. It is an anti- oxidant from marine products and potential for industrial production in future.
基金the supports from the National Natural Science Foundation of China(Nos.31130054 and 31472258)the AoShan Talents Program of Qingdao National Laboratory for Marine Science and Technology(No.2015ASTP-ES02)the Fundamental Research Funds for the Central Universities(No.201564009)
文摘This study applied an optimized one-step 2b-RAD library construction strategy and performed simplified genome sequencing of 539 individuals from three continuously selected Chlamys farreri populations. SNP screening was performed using RAD typing software and population genetic parameters for the continuously selected populations from three generations(G1, G2, G3) were determined. The results showed that the optimized one-step 2b-RAD library construction strategy greatly simplified the experimental process, making it suitable for efficiently constructing a large number of libraries. A total of 18450 SNP markers were identified, which evenly distributed throughout the genome. Population genetic analysis of these three generations showed that the mean value of observed heterozygosity was 0.275 ± 0.177, 0.272 ± 0.181 and 0.275 ± 0.166, respectively. Meanwhile, the mean value of expected heterozygosity was 0.275 ± 0.141, 0.274 ± 0.145 and 0.280 ± 0.133, respectively. The Wright's fixation index(F) was 0.04291, 0.04976 and 0.06685, respectively. Markers deviated from Hardy–Weinberg equilibrium accounted for 10.34%, 12.64%, and 23.11%, and the Shannon diversity index was 0.0999 ± 0.0404, 0.0921 ± 0.0388 and 0.0733 ± 0.0308. FIS(also known as the inbreeding coefficient) of the three populations was 0.0256, 0.0323 and 0.0468, respectively. We suggested that the 2b-RAD method is well suited to population genetic studies of aquacultured organisms. Moreover, our results indicated that the continuous selection affected the population genetic structure of the cultured Penglai-Red scallop, but the change was not significant; therefore, population selection should continue.
基金The National Natural Science Foundation of China under contract Nos 41676147 and 41306117the Scientific and Technological Innovation Project Financially Supported by Qingdao National Laboratory for Marine Science and Technology under contract No.2016ASKJ02+1 种基金the earmarked fund for Modern Agro-industry Technology Research System under contract No.CARS-48the National Basic Research Program of China under contract No.2011CB409805
文摘The biomass and size fraction of phytoplankton in terms of chlorophyll a（Chl a） was measured during four cruises conducted in April, July, October 2013 and January 2014 in mariculture area, the Sanggou Bay, China.Results show that total Chl a levels in the surface seawater of the Sanggou Bay generally range from 0.10 to 20.46μg/L, with an average value of 2.13 μg/L. Nano-phytoplankton was the most important size-fraction and accounted for about 65.1% of total Chl a. In order to evaluate the importance of the ＂protozoan trophic link＂ for energy transfer from the microbial loop to filter-feeding feeders, Zhikong scallop Chlamys farreri was then offered a natural planktonic community as potential prey. Results show that scallops obtained carbon source from natural plankton with the rate of 11 033.05 μg/（g·d）. Protists（nanoflagellates and ciliates） were the dominant source of carbon retained by scallop（48.78%）. The microbial loop provided 58.45% of the carbon source for farmed scallops. These results indicate that the microbial loop represent a valuable trophic resource in mariculture system of the Sanggou Bay.
基金supported by the National Key Research and Development Program of China (No. 2018 YFD0900504)the Qingdao National Laboratory for Marine Science and Technology (No. QNLM2016ORP0307)the Key Technology R&D Program of China (No. 2012BAD17B02)
文摘Bivalve live in aquatic environment and the water temperature can affect their immunity directly.In this research,the scallop Chlamys farreri was injected with 10^4 or 10^7 CFU mL^-1 Vibrio anguillarum and cultured at 11℃,17℃,23℃,and 28℃,respectively.For the control scallop,only phosphate-buffered saline(PBS)was injected.Then total haemocytes and granulocytes were measured by ELISA using monoclonal antibodies.In the meantime,expressions of six immunity-related genes,including lipopolysaccharide andβ-1,3-glucan binding protein(CfLGBP),C-type lectin(CfLec-2),Toll-like receptor(Cf TLR),Lysozyme(CfLYZ),superoxide dismutase(SOD),and phenoloxidase(CfPO)in haemocytes were measured using quantitative real-time PCR.The results showed that total haemocytes counts in 10^4 CFU mL^-1 injection groups showed no differences compared to the control group at all temperatures.However,they varied significantly in 10^7 CFU mL^-1 injection groups at 3 h at 11℃,6–12 h at 17℃,3–48 h at 23℃,and 12–48 h at 28℃.Granulocytes counts in 10^4 CFU mL^-1 injection groups showed no variance compared to the control group at all temperatures,except for 12 h at 23℃,and 24–36 h at 28℃.They were significantly decreased in 10^7 CFU mL^-1 injection groups during 6–48 h at 11℃,12–48 h at 17℃,3–48 h at 23℃,and 3–72 h at 28℃.The expression levels of six immunity-related genes in haemocytes of 10^7 CFU mL^-1 injection groups were significantly higher than those of control group and 10^4 CFU mL^-1 injection groups at all temperatures.The results indicated that infected with high concentration of vibrios,haemocyte counts,granulocyte counts and the expressions of immunity-related genes in scallop C.farreri were significantly affected by environmental temperature.
文摘The effects of membrane penetrating anti-freezing agents(MPAAs), DMSO(dimethyl sulfoxide),glycerol,EG(ethylene glycol) and methanol in combination with different cryoprotective additives suchas carbohydrates,macromolecules and inorganic compounds on the spermatozoon vitality of Chinesescallop,Chlamys farreri,during 1 h 0℃ equilibrium were investigated.When only MPAAs existed,the detrimental effects of different MPAAs ranked in the following order:DMSO【methanol【EG【glycerol.When carbohydrates were added into MPAAs solution,5% glucose caused larger decrease ofspermatozoon vitality than 2.4% lactose.5% glucose or 2.4% lactose in 7.5% glycerol caused com-plete damage.10% yolk was best in maintaining the spermatozoon vitality except when used incombination with 10% methanol.10% milk significantly decreased spermatozoon vitality in EG andmethanol and enhanced its vitality in glycerol,but did not significantly influence it in DMSO.Glycinewas apparently detrimental to spermatozoon vitality.The average
基金Supported by the Shandong Provincial Post-Doctoral Special Fund Innovative Project(No.S43201403)the Post-Doctoral Applied Research Project of Qingdao City(No.Q51201413)the Shandong Provincial Natural Science Foundation,China(No.ZR2015PD004)
文摘Chlamys farreri is an economically important mollusk that can accumulate excessive amounts of cadmium(Cd). Studying the molecular mechanism of Cd accumulation in bivalves is difficult because of the lack of genome background. Transcriptomic analysis based on high-throughput RNA sequencing has been shown to be an efficient and powerful method for the discovery of relevant genes in non-model and genome reference-free organisms. Here, we constructed two c DNA libraries(control and Cd exposure groups) from the digestive gland of C. farreri and compared the transcriptomic data between them. A total of 227 673 transcripts were assembled into 105 071 unigenes, most of which shared high similarity with sequences in the NCBI non-redundant protein database. For functional classification, 24 493 unigenes were assigned to Gene Ontology terms. Additionally, Eu Karyotic Ortholog Groups and Kyoto Encyclopedia of Genes and Genomes analyses assigned 12 028 unigenes to 26 categories and 7 849 unigenes to five pathways, respectively. Comparative transcriptomics analysis identified 3 800 unigenes that were differentially expressed in the Cd-treated group compared with the control group. Among them, genes associated with heavy metal accumulation were screened, including metallothionein, divalent metal transporter, and metal tolerance protein. The functional genes and predicted pathways identified in our study will contribute to a better understanding of the metabolic and immune system in the digestive gland of C. farreri. In addition, the transcriptomic data will provide a comprehensive resource that may contribute to the understanding of molecular mechanisms that respond to marine pollutants in bivalves.
基金This work was supported by the National Science Foundation of China (NO.39970638)and the Science and Technology Bureau of Qingdao (NO.2001-28-50)
文摘To investigate the effect of polypeptide from Chlamys farreri (PCF) on NHDF in vitro, we modeled oxidative damage on normal human dermal fibroblasts (NHDF) exposed to ultraviolet B (UVB). In this study, 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydro-genase (LDH) were tested to measure cell viability. Enzymes including superoxide dismutase (SOD), glu-tathione peroxidase (GSH-PX), catalase (CAT) and xanthine oxidase (XOD) were determined biochemically. Total antioxidative capacity (T-AOC) and anti-superoxide anion capacity (A-SAC) were also determined. Ultrastructure of fibroblasts was observed under transmission electron microscope. The results showed that: UVB (1.176×10-4 J/cm2) suppressed the growth of fibroblasts and the introduction of PCF (0.25%-l%) before UVB reduced the suppression in a concentration-dependent manner. PCF could enhance the activities of SOD, GSH-PX and T-AOC as well as A-SAC. Also PCF could inhibit XOD activity, while it did not affect CAT activity. Ultrastructure of fibroblasts were damaged after UVB irradiation, concentration-dependent PCF reduced the destructive effect of UVB on cells. These results indicated that PCF can protect human dermal fibroblasts from being harmed by UVB irradiation via its antioxidant pro-erty.
